Platelet-Associated Anti-Glycoprotein (GP) IIb-IIIa Autoantibodies in Chronic Immune Thrombocytopenic Purpura Mainly Recognize Cation-Dependent Conformations: Comparison with the Epitopes of Serum Autoantibodies

Satoru Kosugi; Yoshiaki Tomiyama; Masamichi Shiraga; Hirokazu Kashiwagi; Hajime Mizutani; Yuzuru Kanakura; Yoshlyuki Kurata; Yuji Matsuzawa

doi:10.1055/s-0038-1650271

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 1996; 75(02): 339-345
DOI: 10.1055/s-0038-1650271

Original Article

Schattauer GmbH Stuttgart

Platelet-Associated Anti-Glycoprotein (GP) IIb-IIIa Autoantibodies in Chronic Immune Thrombocytopenic Purpura Mainly Recognize Cation-Dependent Conformations: Comparison with the Epitopes of Serum Autoantibodies

Authors

Satoru Kosugi

The Second Department of Internal Medicine, Osaka University Medical School, Osaka, Japan
Yoshiaki Tomiyama

The Second Department of Internal Medicine, Osaka University Medical School, Osaka, Japan
Masamichi Shiraga

The Second Department of Internal Medicine, Osaka University Medical School, Osaka, Japan
Hirokazu Kashiwagi

The Second Department of Internal Medicine, Osaka University Medical School, Osaka, Japan
Hajime Mizutani

The Second Department of Internal Medicine, Osaka University Medical School, Osaka, Japan
Yuzuru Kanakura

The Second Department of Internal Medicine, Osaka University Medical School, Osaka, Japan
Yoshlyuki Kurata

¹Department of Blood Transfusion, Osaka University Hospital, Osaka, Japan
Yuji Matsuzawa

The Second Department of Internal Medicine, Osaka University Medical School, Osaka, Japan

Abstract

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Summary

Platelet-associated and serum anti-glycoprotein (GP) IIb-IIIa autoantibodies were investigated in 57 patients with chronic immune thrombocytopenic purpura (ITP). In modified antigen capture ELISA (MACE) using GPIIb-IIIa-specific-, GPIIb-specific-, and GPIIIa-speci-fic-monoclonal antibody (mAb) for antigen capture, platelet-associated anti-GPIIb-IIIa antibodies were detected in 14 out of 37 patients (38%), and these antibodies could be detected with all 3 mAbs used for antigen capture. In the MACE using EDTA-treated platelets at 37° C, the reactivity of platelet-associated anti-GPIIb-IIIa antibodies in 9 out of 10 patients was markedly reduced in both cases of GPIIb-specific- and GPIIIa-specific-mAb used. Immunoprecipitation experiments further confirmed that the EDTA-treatment abrogated the antigenicity of GPIIb-IIIa for platelet-associated antibodies in 2 patients. Serum anti-GPIIb-IIIa autoantibodies were detected in 23 out of 57 patients (40%). However, only 7 out of 23 serum anti-GPIIb-IIIa antibodies could be detected with all 3 mAbs. The MACE using EDTA-treated platelets further showed that the features of serum anti-GPIIb-IIIa antibodies were different from those of platelet-associated antibodies even in the same patient. Our data demonstrate that the platelet-associated anti-GPIIb-IIIa antibodies mainly recognize the cation-dependent antigenic determinants on GPIIb-IIIa and that serum anti-GPIIb-IIIa antibodies may contain some components which differ in specificity from platelet-associated antibodies.

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Referenzen

References
1 McMillan R. Chronic idiopathic thrombocytopenic purpura. N Engl J Med 1981; 304: 1135-1147
2 Kelton JG, Gibbons S. Autoimmune platelet destruction: idiopathic thrombocytopenic purpura. Semin Thromb Haemost 1982; 8: 83-104
3 McMillan R, Tani P, Millard F, Berchtold P, Renshow L, Woods Jr VL. Platelet-associated and plasma anti-glycoprotein autoantibodies in chronic ITP. Blood 1987; 70: 1040-1045
4 Kiefel V, Santoso S, Kaufmann E, Mueller-Eckhardt C. Autoantibodies against platelet glycoprotein MX: a frequent finding in autoimmune thrombocytopenic purpura. Br J Haematol 1991; 79: 256-262
5 Phillips DR, Charo IF, Parise LV, Fitzgerald LA. The platelet membrane glycoprotein IIb-IIIa complex. Blood 1988; 71: 831-843
6 Tomiyama Y, Kurata Y, Shibata Y, Honda S, Furubayashi T, Mizutani H, Tsubakio T. Immunochemical characterization of an autoantigen on platelet glycoprotein lib in chronic ITP: comparison with the Baka alloantigen. BrJ Haematol 1989; 71: 77-83
7 Kekomaki R, Dawson B, McFarland J, Kunicki TJ. Localization of human platelet autoantigens to the cysteine-rich region of glycoprotein Ilia. J Clin Invest 1991; 88: 847-854
8 Fujisawa K, O’Tool TE, Tani P, Loftus JC, Plow EF, Ginsberg MH, McMillan R. Autoantibodies to presumptive cytoplasmic domain of platelet glycoprotein Ilia in patients with chronic immune thrombocytopenic purpura. Blood 1991; 77: 2207-2213
9 Tomiyama Y, Take H, Honda S, Furubayashi T, Mizutani H, Tsubakio T, Kurata Y. Demonstration of platelet antigens that bind platelet-associated autoantibodies in chronic ITP by direct immunoprecipitation procedure. Br J Haematol 1990; 75: 92-98
10 Berchtold P, Wenger M. Autoantibodies against platelet glycoproteins in autoimmune thrombocytopenic purpura: their clinical significance and response to treatment. Blood 1993; 81: 1246-1250
11 Fujisawa K, Tani P, Piro L, McMillan R. The effect on platelet-associated autoantibody in chronic immune thrombocytopenic purpura. Blood 1993; 81: 2872-2877
12 Fujisawa K, Tani P, McMillan R. Platelet-associated antibody to glycoprotein IIb/IIIa from chronic immune thrombocytopenic purpura patients often binds to divalent cation-dependent antigens. Blood 1993; 81: 1284-1289
13 Kosugi S, Tomiyama Y, Shiraga M, Kashiwagi H, Nakao H, Kanayama Y, Kurata Y, Matsuzawa Y. Cyclic thrombocytopenia associated with IgM anti-GPIIb-IIIa autoantibodies. Br J Haematol 1994; 88: 809-815
14 Tsubakio T, Kurata Y, Yonezawa T, Kitani T. Quantification of platelet-associated IgG with competitive solid-phase enzyme immunoassay. Acta Haematol 1981; 66: 251-256
15 Pidard D, Montgomery RR, Bennett JS, Kunicki TJ. Interaction of AP2, a monoclonal antibody specific for the human platelet glycoprotein IIb-IIIa complex, with intact platelets. J Biol Chem 1983; 258: 12582-12586
16 Newman PJ, Allen RW, Kahn RA, Kunicki TJ. Quantitation of membrane glycoprotein Ilia on intact human platelets using the monoclonal antibody, AP3. Blood 1985; 65: 227-232
17 McEver RR, Benett EM, Martin MN. Identification of two structurally and functionally distinct sites on human platelet membrane glycoprotein IIb-IIIa using monoclonal antibodies. J Biol Chem 1983; 258: 5269-5275
18 Tomiyama Y, Tsubakio T, Piotrowicz RS, Kurata Y, Loftus JC, Kunicki TJ. The Arg-Gly-Asp (RGD) recognition site of platelet glycoprotein IIb-IIIa on nonactivated platelets is accessible to high-affinity macromolecules. Blood 1992; 79: 2303-2312
19 Kuijpers RWAM, Ouwehand WH, Peelen W, Michiels JJ, Engelfriet CP, von dem Borne AEGKr. Thrombocytopenia due to platelet glycoprotein IIb-IIIa-reactive autoantibodies non-reactive with platelets from EDTA blood. Vox Sang 1992; 63: 119-121
20 Kurata Y, Oshida M, Take H, Furubayashi T, Mizutani H, Tomiyama Y, Yonezawa T, Tarui S. Acid treatment of platelets as a simple procedure for distinguishing platelet-specific antibodies from anti-HLA antibodies: comparison with chloroquine treatment. Vox Sang 1990; 59: 106-111
21 Kurata Y, Hayashi S, Kosugi S, Kashiwagi H, Tomiyama Y, Kanayama Y, Matsuzawa Y. Elevated platelet-associated IgG in SLE patients has a nature of anti-platelet autoantibody: differentiation between autoantibodies and immune complexes by ether elution. Br J Haematol 1993; 85: 723-728
22 Pidard D, Didry D, Kunicki TJ, Nurden AT. Temperature-dependent effects of EDTA on the membrane glycoprotein IIb-IIIa complex and platelet aggregability. Blood 1986; 67: 604-611
23 Tomiyama Y, Take H, Ikeda H, Mitani T, Furubayashi T, Mizutani H, Yamamoto N, Tandon NN, Sekiguchi S, Jamieson GA, Kurata Y, Yonezawa T, Tarui S. Identification of the platelet-specific alloantigen, Naka, on platelet membrane glycoprotein IV. Blood 1990; 75: 684-687
24 Kunicki TJ, Newman PJ. The molecular immunology of human platelet proteins. Blood 1992; 80: 1386-1404
25 Newman PJ. Nomenclature of human platelet alloantigens: a problem with HPA system. Blood 1994; 83: 1447-1451
26 Tomiyama Y, Kekomaki R, McFarland J, Kunicki TJ. Antivinculin antibodies in sera of patients with immune thrombocytopenia and in sera of normal subjects. Blood 1992; 79: 161-168